title: Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the regulatory domain of aspartokinase (Rv3709c) from Mycobacterium tuberculosis.
abstract: The regulatory domain of Mycobacterium tuberculosis aspartokinase (Mtb-AK, Mtb-Ask, Rv3709c) has been cloned, heterologously expressed in Escherichia coli and purified using standard chromatographic techniques. Screening for initial crystallization conditions using the regulatory domain (AK-β) in the presence of the potential feedback inhibitor threonine identified four conditions which yielded crystals suitable for X-ray diffraction analysis. From these four conditions five different crystal forms of Mtb-AK-β resulted, three of which belonged to the orthorhombic system, one to the tetragonal system and one to the monoclinic system. The highest resolution (1.6 Å) was observed for a crystal form belonging to space group P2(1)2(1)2(1), with unit-cell parameters a=53.70, b=63.43, c=108.85 Å and two molecules per asymmetric unit.
authors.affiliation: EMBL Hamburg Outstation, c/o DESY, Notkestrasse 85, D-22603 Hamburg, Germany. lschuldt@mb.au.dk
authors.forename: Linda
authors.initials: L
authors.lastname: Schuldt
authors.forename: Ruth
authors.initials: R
authors.lastname: Suchowersky
authors.forename: Katharina
authors.initials: K
authors.lastname: Veith
authors.forename: Jochen
authors.initials: J
authors.lastname: Mueller-Dieckmann
authors.forename: Manfred S
authors.initials: MS
authors.lastname: Weiss
mesh_terms: D001222:Aspartate Kinase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D001222:Aspartate Kinase